PITX3 Methylation Analysis Protocol

For the analysis of PITX3 methylation, the FFPET samples were processed according to the InnuCONVERT Bisulfite All-In-One Kit (Analytik Jena, Germany) as previously published [10 (link)]. To validate the assay performance, a serial dilution of bisulfite-converted artificially methylated DNA (CpGenome™ Universal Methylated DNA; Merck Millipore, Darmstadt, Germany) and unmethylated DNA from human sperm (NW Andrology & Cryobank Inc., Spokane, WA, USA) was used. As a calibrator sample DNA, a 1:1 mixture of bisulfite-converted unmethylated and artificially methylated DNA was used.

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Holmes E.E., Goltz D., Sailer V., Jung M., Meller S., Uhl B., Dietrich J., Röhler M., Ellinger J., Kristiansen G, & Dietrich D. (2016). PITX3 promoter methylation is a prognostic biomarker for biochemical recurrence-free survival in prostate cancer patients after radical prostatectomy. Clinical Epigenetics, 8, 104.

Publication 2016

InnuCONVERT Bisulfite All-In-One Kit CpGenome™ Universal Methylated DNA

Assay Bisulfite Dilution Human Sperm

Corresponding Organization :

Other organizations : University Hospital Bonn, Cornell University

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Variable analysis

independent variables

Bisulfite-converted artificially methylated DNA (CpGenome™ Universal Methylated DNA)
Unmethylated DNA from human sperm (NW Andrology & Cryobank Inc., Spokane, WA, USA)

dependent variables

PITX3 methylation

control variables

Bisulfite conversion process using the InnuCONVERT Bisulfite All-In-One Kit

controls

Positive control: 1:1 mixture of bisulfite-converted unmethylated and artificially methylated DNA as a calibrator sample
Negative control: Not explicitly mentioned

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